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BACKGROUND: Syndrome coronavirus-2 (SARS-CoV-2) has developed various strategies to evade the antiviral impact of type I IFN. Non-structural proteins and auxiliary proteins have been extensively researched on their role in immune escape. Nevertheless, the detailed mechanisms of structural protein-induced immune evasion have not been well elucidated. METHODS: Human alveolar basal epithelial carcinoma cell line (A549) was stimulated with polyinosinic-polycytidylic acid (PIC) and independently transfected with four structural proteins expression plasmids, including nucleocapsid (N), spike (S), membrane (M) and envelope (E) proteins. By RT-qPCR and ELISA, the structural protein with the most pronounced inhibitory effects on IFN-ß induction was screened. RNA-sequencing (RNA-Seq) and two differential analysis strategies were used to obtain differentially expressed genes associated with N protein inhibition of IFN-ß induction. Based on DIANA-LncBase and StarBase databases, the interactive competitive endogenous RNA (ceRNA) network for N protein-associated genes was constructed. By combining single-cell sequencing data (GSE158055), lncRNA-miRNA-mRNA axis was further determined. Finally, RT-qPCR was utilized to illustrate the regulatory functions among components of the ceRNA axis. RESULTS: SARS-CoV-2 N protein inhibited IFN-ß induction in human alveolar epithelial cells most significantly compared with other structural proteins. RNA-Seq data analysis revealed genes related to N protein inhibiting IFNs induction. The obtained 858 differentially expressed genes formed the reliable ceRNA network. The function of LINC01002-miR-4324-FRMD8 axis in the IFN-dominated immune evasion was further demonstrated through integrating single-cell sequencing data. Moreover, we validated that N protein could reverse the effect of PIC on LINC01002, FRMD8 and miR-4324 expression, and subsequently on IFN-ß expression level. And LINC01002 could regulate the production of FRMD8 by inhibiting miR-4324. CONCLUSION: SARS-CoV-2 N protein suppressed the induction of IFN-ß by regulating LINC01002 which was as a ceRNA, sponging miR-4324 and participating in the regulation of FRMD8 mRNA. Our discovery provides new insights into early intervention therapy and drug development on SARS-CoV-2 infection.
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COVID-19 , MicroRNAs , RNA Longo não Codificante , Humanos , Células A549 , Proteínas do Nucleocapsídeo de Coronavírus/genética , Proteínas do Nucleocapsídeo de Coronavírus/metabolismo , COVID-19/virologia , COVID-19/imunologia , Evasão da Resposta Imune , Interferon beta/genética , Interferon beta/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Fosfoproteínas , RNA Endógeno Competitivo , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismoRESUMO
BACKGROUND: Nosocomial infections with heavy disease burden are becoming a major threat to the health care system around the world. Through long-term, systematic, continuous data collection and analysis, Nosocomial infection surveillance (NIS) systems are constructed in each hospital; while these data are only used as real-time surveillance but fail to realize the prediction and early warning function. Study is to screen effective predictors from the routine NIS data, through integrating the multiple risk factors and Machine learning (ML) methods, and eventually realize the trend prediction and risk threshold of Incidence of Nosocomial infection (INI). METHODS: We selected two representative hospitals in southern and northern China, and collected NIS data from 2014 to 2021. Thirty-nine factors including hospital operation volume, nosocomial infection, antibacterial drug use and outdoor temperature data, etc. Five ML methods were used to fit the INI prediction model respectively, and to evaluate and compare their performance. RESULTS: Compared with other models, Random Forest showed the best performance (5-fold AUC = 0.983) in both hospitals, followed by Support Vector Machine. Among all the factors, 12 indicators were significantly different between high-risk and low-risk groups for INI (P < 0.05). After screening the effective predictors through importance analysis, prediction model of the time trend was successfully constructed (R2 = 0.473 and 0.780, BIC = -1.537 and -0.731). CONCLUSIONS: The number of surgeries, antibiotics use density, critical disease rate and unreasonable prescription rate and other key indicators could be fitted to be the threshold predictions of INI and quantitative early warning.
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Infecção Hospitalar , Aprendizado de Máquina , Humanos , Infecção Hospitalar/epidemiologia , Medição de Risco/métodos , China/epidemiologia , Fatores de Risco , IncidênciaRESUMO
BACKGROUND: As a histone methyltransferase, suppressor of variegation 3-9 homolog 1 (SUV39H1) plays an important role in the occurrence and development of cancer. To explore the mechanism and biological function of SUV39H1 in hepatitis B virus-associated hepatocellular carcinoma (HBV-HCC) can gain an insight into the pathogenesis of HBV-HCC. METHODS: The effect of HBV infection on SUV39H1 in hepatoma cells was detected. CCK-8, colony growth assay and wound healing assay were used to assess the proliferation and migration of HBV-positive hepatoma cells. RNA sequencing (RNA-seq) was applied to find differential genes and enriched pathways. The serum SUV39H1 level in HBV-HCC patients was detected and its correlation with clinical indicators was analyzed. RESULTS: SUV39H1 was increased by HBV infection and promoted the proliferation and migration of hepatoma cells. SUV39H1 could upregulate the expression of mitochondrial oxidative phosphorylation (OXPHOS) pathway-related genes. OXPHOS pathway inhibitors could reduce the capacity of proliferation and migration of hepatoma cells after overexpressing SUV39H1. Serum SUV39H1 levels were higher in chronic hepatitis B (CHB) patients than in healthy controls and higher in HBV-HCC patients than in CHB patients. In the diagnosis of HCC, the predictive value of SUV39H1 combined with alpha-fetoprotein (AFP) was better than that of AFP alone. CONCLUSION: SUV39H1 is regulated by HBV infection and promotes the proliferation and migration of hepatoma cells by targeting OXPHOS pathway. It indicates that SUV39H1 may be a new biomarker of the diagnosis of HCC.
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Carcinoma Hepatocelular , Hepatite B Crônica , Hepatite B , Neoplasias Hepáticas , Humanos , Carcinoma Hepatocelular/patologia , Vírus da Hepatite B/metabolismo , alfa-Fetoproteínas/metabolismo , Neoplasias Hepáticas/patologia , Fosforilação Oxidativa , Biomarcadores , Hepatite B/complicações , Hepatite B Crônica/complicações , Hepatite B Crônica/patologia , Metiltransferases/metabolismo , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismoRESUMO
Group B Streptococcus (GBS) colonizes the gastrointestinal and urogenital tracts of approximately 30% of women, and it can cause sepsis and meningitis in neonates. GBS has been shown to form biofilms in vitro, but the effects of environmental and genotypic factors upon GBS biofilm formation are unclear. The aim of the present study was to optimize culture conditions for enhanced GBS biofilm production. Furthermore, this study also investigated the influences of strain lineage, pilus profile, and isolation source on GBS biofilm formation. The results demonstrate that the fed-batch mode and acidic pH strongly enhanced GBS biofilm formation in vitro. These findings suggest that the fed-batch mode may be suitable for both screening and fundamental studies of GBS biofilm formation. Moreover, this study demonstrated a correlation between the hyper virulent clonal complex 17 and a strong biofilm phenotype.
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Biofilmes/crescimento & desenvolvimento , Streptococcus agalactiae/genética , Técnicas de Cultura Celular por Lotes , Fímbrias Bacterianas/genética , Fímbrias Bacterianas/metabolismo , Genótipo , Concentração de Íons de Hidrogênio , Modelos Logísticos , Fenótipo , Streptococcus agalactiae/crescimento & desenvolvimentoRESUMO
We explored the use of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for identification of Fusobacterium nucleatum subspecies. MALDI-TOF MS spectra of five F. nucleatum subspecies (animalis, fusiforme, nucleatum, polymorphum, and vincentii) were analyzed and divided into four distinct clusters, including subsp. animalis, nucleatum, polymorphum, and fusiforme/vincentii. MALDI-TOF MS with the modified SARAMIS database further correctly identified 28 of 34 F. nucleatum clinical isolates to the subspecies level.
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Técnicas de Tipagem Bacteriana , Fusobacterium nucleatum/classificação , Fusobacterium nucleatum/genética , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Infecções por Fusobacterium/microbiologia , Humanos , RNA Bacteriano/genética , RNA Ribossômico 16S/genéticaRESUMO
Rapid and accurate diagnosis of influenza is important for infection control, as well as for patient management. Alere i Influenza A&B is an isothermal nucleic acid amplification-based integrated system for detection and differentiation of influenza virus A and influenza virus B. The performance of the Alere i Influenza A&B was screened using frozen nasopharyngeal-swab specimens collected in viral transport medium (VTM) that were originally tested fresh with the FilmArray Respiratory Panel (RP) assay during the 2012-2013 influenza outbreak. In total, 360 VTM specimens were selected for Alere i Influenza A&B testing: 40 influenza virus A H1N1-2009 (influenza virus A-1), 40 influenza virus A H3N2 (influenza virus A-3), 37 influenza virus A "equivocal" or "no subtype detected" (influenza virus A-u), 41 influenza virus B, and 202 influenza virus-negative specimens, as initially determined by the FilmArray RP assay. The Alere assay showed sensitivities of 87.2%, 92.5%, 25.0%, and 97.4% for influenza virus A-1, influenza virus A-3, influenza virus A-u, and influenza virus B, respectively, after discordant resolution by Prodesse ProFLU+ PCR. The specificities were 100% for both influenza virus A and influenza virus B. In general, the Alere i Influenza A&B provided good sensitivity, although the assay did show poorer sensitivity with samples determined to have low influenza virus A titers by Prodesse ProFlu+ PCR (a mean real-time PCR threshold cycle [CT] value of 31.9 ± 2.0), which included the majority of the samples called influenza virus A "equivocal" or "no subtype detected" by a single BioFire FilmArray RP test. The integrated, rapid, and simple characteristics of the Alere i Influenza A&B assay make it a potential candidate for point-of-care testing, with a test turnaround time of less than 15 min.
Assuntos
Vírus da Influenza A/isolamento & purificação , Vírus da Influenza B/isolamento & purificação , Influenza Humana/diagnóstico , Influenza Humana/virologia , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Nasofaringe/virologia , Sistemas Automatizados de Assistência Junto ao Leito , Estudos Retrospectivos , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: The accumulated evidence demonstrates that stress plays an important role in the pathogenesis of depression that is associated with intestinal dysfunctions. However, the mechanisms remain unresolved. METHODS: A total of 40 male Wistar rats were obtained and randomly divided into four equal-sized group: control, PDTC + chronic and unpredictable mild stress (CUMS), FLX + CUMS, and CUMS. Western blotting and qRT-PCR were used to examine the levels of nitric oxide (NO), nuclear factor kappa beta (NF-κB), inducible nitric oxide synthase (iNOS), and iNOS mRNA in spinal cord L1-2 and colon. KEY RESULTS: Chronic and unpredictable mild stress increased the serum CORT level, decreased body weight and sucrose preference, and altered OFT performance, while increased levels of NO, iNOS mRNA, iNOS and NF-κB protein in colon and spinal cord were accompanied by histopathological changes in colon. Pretreatment with an NF-κB inhibitor, pyrrolidine dithiocarbamate (PDTC), reversed these effects. Fluoxetine failed to prevent NO increase in both spinal cord and colon, while the iNOS protein level, although not statistically significantly increased compared to control, was not decreased compared to CUMS. Also, fluoxetine failed to prevent histological changes. CONCLUSION: In conclusion, the NF-κB/iNOS pathway may be involved in the mechanism of CUMS-induced depressive-like behavior and colon tissue injury.
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Colorization for fabricating aluminum pigments has broad application prospects in recent years. In this study, yellow-colored aluminum pigments with the double-layer structure Al@SiO2@PFMV were prepared using a sol-gel method. A crosslinked copolymeric dye (PFMV) was firstly synthesized by radical polymerization using vinyl triethoxysilane (VTES) and a small molecular dye (FGMAC) as monomers. Then, colored aluminum pigments were prepared by hydrolysis and condensation of the copolymers on the surface of aluminum pigments. SEM, AFM, FTIR, and XPS were used to characterize the surface morphology and chemical structure of the colored aluminum pigments. It was found that the colored aluminum pigments have a heterogeneous and smooth surface layer. The anticorrosion results showed that the colored aluminum pigments had better chemical stability with significantly improving corrosion resistance compared to raw aluminum pigments and Al@SiO2 with the single-layer coating. Chromatism analysis indicated that the lightness of Al@SiO2@PFMV pigments decreased slightly and the color changed from silver-gray to yellow.
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Group B streptococcus (GBS) is a leading cause of invasive neonatal infections and has increasingly been associated with invasive diseases in non-pregnant adults. We collected 113 GBS isolates recovered from sterile and non-sterile specimens from seven tertiary hospitals in China between October 2014 and September 2016. Medical records were retrospectively reviewed and the sequence types, serotypes, virulence, and antimicrobial resistance profiles of the isolates were characterized and correlated. Significantly higher C-reactive protein and procalcitonin levels and absolute neutrophil counts were observed in patients with invasive infections than in those with non-invasive infections (Pâ¯<â¯0.05). The 113 isolates were grouped into 24 sequence types, 5 clonal complexes, and 6 serotypes. multivariate analysis revealed that clonal complex 17 isolates characterized by serotype iii, the surface protein gene rib, and the pilus island pi-2b were independently correlated with invasive infection (or: 6.79; 95% ci: 2.31-19.94, Pâ¯<â¯0.001). These results suggest alternative molecular biomarkers for diagnosis and prognosis of GBS infections.
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Infecções Estreptocócicas/microbiologia , Streptococcus/isolamento & purificação , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/uso terapêutico , Criança , Pré-Escolar , China , Farmacorresistência Bacteriana/efeitos dos fármacos , Farmacorresistência Bacteriana/genética , Feminino , Genótipo , Humanos , Lactente , Recém-Nascido , Masculino , Testes de Sensibilidade Microbiana/métodos , Pessoa de Meia-Idade , Tipagem de Sequências Multilocus/métodos , Estudos Retrospectivos , Sorogrupo , Sorotipagem/métodos , Infecções Estreptocócicas/tratamento farmacológico , Streptococcus/efeitos dos fármacos , Streptococcus/genética , Adulto JovemRESUMO
Previous results regarding the associations between esophageal squamous-cell carcinoma (ESCC) risk and smoking/alcohol drinking in high-risk areas are inconsistent. We performed a large population-based case-control study from 2010 to 2013 in a high-incidence area of China, and enrolled 1353 ESCC cases and 1961 controls. Data regarding smoking and alcohol drinking were collected via face-to-face interviews using a structured questionnaire. Odd ratios (ORs) with 95% confidence intervals (CIs) were estimated using unconditional logistic regression models. After adjusting for alcohol drinking and other potential confounders, male heavy smokers (i.e., those who started smoked more than 20 cigarettes per day or 40 pack-years, or started smoking early), showed a moderately increased risk for ESCC; however, current smoking was not associated with an increased risk. Alcohol drinking among males significantly increased the risk for ESCC (OR = 2.20, 95%CI:1.79~2.70). We observed increasing excess ESCC risks with decreasing age at behavior initiation as well as with increasing duration and intensity of alcohol intake, which were particularly evident among current smokers. In contrast, neither smoking nor alcohol drinking was not associated with ESCC risk among females. In conclusion, alcohol drinking shows a monotonic dose-response relationship with ESCC risk among men, and this relationship is particularly evident among smokers.
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Consumo de Bebidas Alcoólicas/efeitos adversos , Carcinoma de Células Escamosas do Esôfago/epidemiologia , Fumar/efeitos adversos , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , China/epidemiologia , Carcinoma de Células Escamosas do Esôfago/etiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , RiscoRESUMO
Radical prostatectomy (RP) is the gold standard for the treatment of localized PCa. A meta-analysis was conducted to evaluate the effect of different techniques of pelvic floor reconstruction on urinary continence. A comprehensive search was made for trials that evaluated the efficacy of pelvic floor reconstruction. Relevant databases included PubMed, Embase, Cochrane, Ovid, Web of Science databases and relevant trials from the references. Random-effects model was used to estimate risk ratios (RRs) statistics. Pooled results of patients treated with posterior reconstruction (PR) demonstrated complete urinary continence improved at 1-4, 28-42, 90, 180 and 360 days following catheter removal. Anterior suspension (AS) was associated with improvement only at 28-42 days. The anterior reconstruction (AR) + PR was associated with urinary continence at 1-4, 90 and 180 days. AS + PR was not associated with any benefit. And PR improved social urinary continence at 7-14 and 28-42 days. No benefit was associated with AS. AR + PR had better outcomes at 90 and 180 days. AS + PR was significant improved at 28-42 and 90 days. Patients who underwent RP and PR had the least urinary incontinence. No significant benefit was observed after AS. AR + PR and AS + PR had little benefit in the post-operative period.
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Diafragma da Pelve/cirurgia , Procedimentos de Cirurgia Plástica , Cuidados Pós-Operatórios , Prostatectomia , Ensaios Clínicos como Assunto , Estudos de Coortes , Humanos , Masculino , Razão de Chances , Cuidados Pós-Operatórios/métodos , Prostatectomia/efeitos adversos , Viés de Publicação , Procedimentos de Cirurgia Plástica/métodos , Resultado do Tratamento , Incontinência UrináriaRESUMO
Human plasminogen activator inhibitor-1 (PAI-1) is closely related to embryonic development and pregnancy success. The association between PAI-1 gene polymorphisms (PAI-1-844G/A and PAI-1-675G/A) and the risk of recurrent pregnancy loss (RPL) is controversial. Therefore, we perform this review to clarify the association between PAI-1 gene polymorphisms and RPL risk. We performed a systematic search for studies that described the effect of PAI-1 polymorphisms on RPL risk. The odds ratios (ORs) with corresponding 95% confidence intervals (CIs) were considered under recessive genetic models. Furthermore, we conducted a subgroup analysis based on the studies' geographic regions of origin. Data were analyzed using Stata 11.2 software. Eighteen studies were included, and a high degree of statistical heterogeneity existed among the studies. In this study, we found a significant association between the PAI-1-675G/A polymorphism and the risk of RPL under the recessive model (OR = 1.70, 95% CI = 1.21-2.38). However, no significant association between the PAI-1-844G/A polymorphism and RPL was noted. PAI-1-675G/A (4G/5G) polymorphisms play a potential role in RPL. The screening of PAI-1 (4G/5G) gene mutations should be included during an RPL diagnostic workup, and patients should be treated using anticoagulant therapy during pregnancy if necessary.
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Perda do Embrião/genética , Predisposição Genética para Doença/genética , Inibidor 1 de Ativador de Plasminogênio/genética , Polimorfismo Genético/genética , Feminino , Humanos , Gravidez , Fatores de RiscoRESUMO
Esophageal carcinoma (EC) is a serious malignancy, and its epidemiologic etiology is not fully explained. We performed this review to investigate the association between teeth loss and teeth brushing and the risk of EC. A systematic search was conducted to identify all relevant studies. The Q test and I(2) statistic were used to examine between-study heterogeneity. Pooled odds ratios (ORs) with corresponding 95% confidence intervals (CIs) were considered by fixed or random effects models. Furthermore, we conducted subgroup analyses based on study design, the studies' geographic regions and case type of origin. Modified Egger linear regression test was used to estimate publication bias. Ten articles were included. Pooled analyses indicated that teeth loss was associated with an increased risk of EC for Asians (OR, 1.52; 95% CI: 1.30, 1.78), and high frequency of teeth brushing was associated with a lower incidence of EC (OR, 0.62; 95%CI: 0.43, 0.89). Subgroup analyses showed consistent results and no publication bias existed. Teeth loss and teeth brushing play potential roles in the progressing of EC. People should take care of their oral health in daily life. And large well-designed researches are needed to fully describe the association between teeth health and EC risk.
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Neoplasias Esofágicas/diagnóstico , Neoplasias Esofágicas/epidemiologia , Perda de Dente/epidemiologia , Escovação Dentária/estatística & dados numéricos , Adolescente , Adulto , Distribuição por Idade , Idoso , Idoso de 80 Anos ou mais , Causalidade , Criança , Pré-Escolar , Comorbidade , Feminino , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Prognóstico , Medição de Risco/métodos , Distribuição por Sexo , Perda de Dente/diagnóstico , Perda de Dente/prevenção & controle , Adulto JovemRESUMO
OBJECTIVES: Several preclinical and observational studies have shown that anti-diabetic medications (ADMs) may modify the risk of lung cancer. We performed a systematic review and meta-analysis evaluating the effect of metformin, sulfonylureas (SUs), thiazolidinediones (TZDs), and insulin on the risk of lung cancer in patients with diabetes mellitus (DM). MATERIALS AND METHODS: We conducted a systematic search of Pubmed and Web of Science, up to August 20, 2013. We also searched the Conference Proceedings Citation Index (CPCI) and China National Knowledge Infrastructure (CNKI) for abstracts from major meetings. Fixed or random effect pooled measures were selected based on heterogeneity among studies, which was evaluated using Q test and the I2 of Higgins and Thompson. Meta-regression was used to explore the sources of between-study heterogeneity. Publication bias was analyzed by Begg's funnel plot and Egger's regression test. Associations were assessed by odds ratios (ORs) with 95% confidence intervals (CIs). RESULTS: A total of 15 studies (11 cohort, 4 case-control) were included in this meta-analysis. In observational studies no significant association between metformin (n=11 studies; adjusted OR=0.99, 95%CI: 0.87-1.12), SUs (n=5 studies; adjusted OR=0.98, 95%CI: 0.79-1.22), or TZDs (n=7 studies; adjusted OR=0.92, 95%CI: 0.75-1.13), insulin (n=6 studies; adjusted OR=1.13, 95%CI: 0.79-1.62) use and risk of developing lung cancer was noted. There was considerable inherent heterogeneity between studies not explained by study design, setting, or location. CONCLUSIONS: Meta-analysis of existing studies does not support a protective or harmful association between ADMs use and risk of lung cancer in patients with DM. There was considerable heterogeneity across studies, and future, well-designed, prospective studies would be required for better understanding of any association.
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Diabetes Mellitus/tratamento farmacológico , Hipoglicemiantes/efeitos adversos , Hipoglicemiantes/uso terapêutico , Neoplasias Pulmonares/epidemiologia , Humanos , Insulina/efeitos adversos , Insulina/uso terapêutico , Metformina/efeitos adversos , Metformina/uso terapêutico , Compostos de Sulfonilureia/efeitos adversos , Compostos de Sulfonilureia/uso terapêutico , Tiazolidinedionas/efeitos adversos , Tiazolidinedionas/uso terapêuticoRESUMO
PURPOSE: Aiming to identify macrolide and beta-lactam resistance in clinical bacterial isolates rapidly and accurately, a two-step algorithm was developed based on detection of eight antibiotic resistance genes. METHODS: Targeting at genes linked to bacterial macrolide (msrA, ermA, ermB, and ermC) and beta-lactam (blaTEM, blaSHV, blaCTX-M-1, blaCTX-M-9) antibiotic resistances, this method includes a multiplex real-time PCR, a melting temperature profile analysis as well as a liquid bead microarray assay. Liquid bead microarray assay is applied only when indistinguishable Tm profile is observed. RESULTS: The clinical validity of this method was assessed on clinical bacterial isolates. Among the total 580 isolates that were determined by our diagnostic method, 75% of them were identified by the multiplex real-time PCR with melting temperature analysis alone, while the remaining 25% required both multiplex real-time PCR with melting temperature analysis and liquid bead microarray assay for identification. Compared with the traditional phenotypic antibiotic susceptibility test, an overall agreement of 81.2% (kappa=0.614, 95% CI=0.550-0.679) was observed, with a sensitivity and specificity of 87.7% and 73% respectively. Besides, the average test turnaround time is 3.9h, which is much shorter in comparison with more than 24h for the traditional phenotypic tests. CONCLUSIONS: Having the advantages of the shorter operating time and comparable high sensitivity and specificity with the traditional phenotypic test, our two-step algorithm provides an efficient tool for rapid determination of macrolide and beta-lactam antibiotic resistances in clinical bacterial isolates.